Therapeutics for thrombocytopenia

ABSTRACT

Compounds represented by the general formula (I) or pharmacologically acceptable salts thereof: ##STR1## (where R is a group --NHCHR 1  R 2 , --N(CHR 1  R 2 ) 2 , --N(CHR 1  R 2 )CHR 3  R 4 , --N +  (CHR 1  R 2 ) 3 , --N +  (CHR 1  R 2 ) 2  CHR 3  R 4  or --N +  (CHR 1  R 2 )(CHR 3  R 4 )CHR 5  R 6  (where R 1 , R 2 , R 3 , R 4 , R 5  and R 6 , which may be the same or different, are each a hydrogen atom, an alkyl group, an alkenyl group, an aryl group or an aralkyl group, or an alkyl, alkenyl, aryl or aralkyl group which is substituted by at least one substituent selected from the group consisting of a halogen atom, a hydroxyl group, a lower alkoxy group, a lower alkylacyloxy group, a lower alkylacyl group, a lower alkoxycarbonyl group, a nitro group, a cyano group and a heterocyclic group, with CHR 1  R 2 , CHR 3  R 4  or CHR 5  R 6  optionally forming a cyclic alkyl group, provided that when R is --N(CHR 1  R 2 )CHR 3  R 4 , --N +  (CHR 1  R 2 ) 2  CHR 3  R 4  or --N +  (CHR 1  R 2 )(CHR 3  R 4 )CHR 5  R 6 , CHR 1  R 2 , CHR 3  R 4  and CHR 5  R 6  are different groups); therapeutics for thrombocytopenia containing them as an effective ingredient; as well as intermediates for their synthesis and processes for producing them.

This application is the national phase of international applicationPCT/JP96/03008, filed Oct. 17, 1996 which designated the U.S. now WO97/14704 Apr. 24, 1997.

TECHNICAL FIELD

This invention relates to novel 2-pyranone derivatives andpharmacologically acceptable salts thereof, as well as therapeutics forthrombocytopenia containing them as an effective ingredient. Theinvention also relates to intermediates for their synthesis andprocesses for producing them.

BACKGROUND ART

Thrombocytopenia is a disease that accompanies immunological disordersor bone marrow damaging metastatic tumors, tuberculosis, leukemia, etc.Alternatively, it is caused by other factors such as the use ofchemotherapeutics or radiation therapy. Thrombocytopenia is a seriousdisease which, when aggravated, causes bleeding in various parts of thebody, occasionally leading to death.

Symptomatic therapy by platelet transfusion is currently considered tobe the sole reliable method that can treat thrombocytopenia and it isdesired to develop therapeutics that can increase platelets per se.

In recent years, reports have been made that show the plateletincreasing action of cytokines such as interleukin-6, interleukin-11 andleukemia inhibitory factor (LIF) (Ishibashi et al., Blood, 74:1241-1244,1989; Asano et al., Blood, 75:1602-1605, 1990; Zenji Okada et al.,KETUEKI SHUYOKA, 22:23-31, 1991). However, the production of thesecytokines is regulated and controlled by various cells within the bodyand if they are externally administered to the body, the balance inregulation is upset, eventually causing serious side effects such asdamage to the liver.

It has also been suggested recently that a protein called"thrombopoietin (TPO)" is a factor that increases megakaryocytes andplatelets (see, for example, Sauvage et al., Nature, 369:533-538, 1994),however, clinical effects of this protein have not yet been verified.

Derivatives such as muramyl dipeptide are known as low-molecular weightcompounds that increase the platelet count (Nakajima et al.,Arzneim.-Forsch./Drug Res. 41:60-65, 1989). It is postulated that thesederivatives increase platelets by activating monocytes and macrophagesso as to produce interleukin-6. However, it has also been reported thatthe administration of derivatives such as muramyl dipeptide alsotriggers other physiological activities based on the activation ofmacrophages, thereby causing fever and other side effects (NIHON IGAKUHOSHASEN GAKKAISHI, 48(4):514, 1988).

The compounds structurally similar to the compounds of the invention aretaught in Japanese Patent Public Disclosure Nos. 304893/1989 and186/1990, as well as The Journal of Antibiotics, 42:1331-1343, 1989;they are 2-pyranone derivatives obtained as the metabolites ofactinomyces of the genus Streptomyces and these compounds have beenreported to have an antimicrobial action against plant pathogenic fungi,as well as cytotoxicity to leukemic cells.

In addition, Japanese Patent Public Disclosure Nos. 213758/1993 and2886/1995 teach that 2-pyranone derivatives are compounds exhibiting aplatelet increasing action in mouse. However, these compounds are notnecessarily satisfactory in terms of safety.

An object of the invention is to overcome the aforementioned defects ofthe prior art by providing compounds that are safe and which have anaction for increasing platelets per se.

DISCLOSURE OF INVENTION

The present inventors conducted intensive studies on various compoundswith a view to solving the aforementioned problems and found novel2-pyranone derivatives that had a platelet increasing action in mouseand which yet had low toxicity. The present invention has beenaccomplished on the basis of this finding.

Thus, the present invention provides compounds represented by thegeneral formula (I) or pharmacologically acceptable salts thereof:##STR2## (where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂, --N(CHR₁R₂)CHR₃ R₄, --N⁺ (CHR₁ R₂)₃, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁R₂)(CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃, R₄, R₅ and R₆, which may be thesame or different, are each a hydrogen atom, an alkyl group, an alkenylgroup, an aryl group or an aralkyl group, or an alkyl, alkenyl, aryl oraralkyl group which is substituted by at least one substituent selectedfrom the group consisting of a halogen atom, a hydroxyl group, a loweralkoxy group, a lower alkylacyloxy group, a lower alkylacyl group, alower alkoxycarbonyl group, a nitro group, a cyano group and aheterocyclic group, with CHR₁ R₂, CHR₃ R₄ or CHR₅ R₆ optionally forminga cyclic alkyl group, provided that when R is --N(CHR₁ R₂)CHR₃ R₄, --N⁺(CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁ R₂)(CHR₃ R₄)CHR₅ R₆, CHR₁ R₂, CHR₃ R₄,and CHR₅ R₆ are different groups)).

The invention also provides therapeutics for thrombocytopenia thatcontain those compounds or pharmcologically acceptable salts thereof, aswell as intermediates for their synthesis and processes for producingthem.

With the 2-pyranone derivatives (I) of the invention, preferred examplesof R₁, R₂, R₃, R₄, R₅ and R₆ include the following: exemplary alkylgroups are straight-chained or branched lower alkyl groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, hexyl and octylgroups; exemplary alkenyl groups are straight-chained or branched loweralkenyl groups such as vinyl, allyl, 1-propenyl, 2-butenyl and2-methyl-2-propenyl groups; exemplary aryl groups are C₆₋₁₀ aryl groupssuch as phenyl, tolyl, xylyl, mesityl, cumenyl and naphtyl groups; andexemplary aralkyl groups are C₇₋₂₄ aralkyl groups such as benzyl,phenethyl, trityl and benzhydryl groups. Examples of the cyclic alkylgroup formed by CHR₁ R₂, CHR₃ R₄ or CHR₅ R₆ include mono- or polycyclicalkyl groups such as cyclopentyl, cyclohexyl, cycloheptyl, menthyl,phentyl and bornyl groups. Unless otherwise noted, the term "lower" asused herein means preferably 1-8 carbon atoms, with the range of 1-4carbon atoms being particularly preferred.

The above-defined alkyl, alkenyl, aryl and aralkyl groups may besubstituted with the following: halogen atoms such as fluorine, chlorineand bromine atoms; a hydroxyl group; lower alkoxy groups such asmethoxy, ethoxy and propoxy groups; lower alkylacyloxy groups such asacetoxy and propionyloxy groups; lower alkylacyl groups such as acetyl,propionyl and butyryl groups; lower alkoxycarbonyl groups such asmethoxycarbonyl, ethoxycarbonyl and propoxycarbonyl groups; a nitrogroup, a cyano group; and heterocyclic groups which are preferablyunsaturated mono-heterocyclic groups such as pyrrolyl, pyridyl,pyrazolyl, imidazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazolyl(e.g., 4H-1,2,4-triazolyl), tetrazolyl (e.g., 1H-tetrazolyl or2H-tetrazolyl), furyl, thiophenyl, benzofuranyl, benzothiophenyl,oxazolyl, isoxazolyl, thiazolyl, indolyl, benzothiazolyl, benzimidazolyland quinolyl. These heterocyclic groups may be bound by carbon atoms or,alternatively, they may be bound by nitrogen atoms to formintramolecular quaternary salts.

The 2-pyranone derivatives (I) of the invention may be used in the formof pharmacologically acceptable nontoxic salts with inorganic metalssuch as alkali metals (e.g. sodium and potassium) and alkaline earthmetals (e.g. calcium and magnesium); basic amino acids such as lysineand arginine; and organic amines such as ammonium.

Further, the 2-pyranone derivatives (I) of the invention can be used inthe form pharmacologically acceptable nontoxic acid addition salts. Suchacid addition salts include, but are not limited to, inorganic acidsalts such as hydrochlorides, sulfates, hydrobromides and phosphates;organic acid salts such as formates, acetates, succinates, maleates,fumarates, malates, mandelates, glutamates, aspartates,methanesulfonates and p-toluenesulfonates.

The 2-pyranone derivatives (I) of the invention and the compoundsrepresented by the general formula (II) to be set forth below which areused as intermediates for synthesis of those derivatives have variousisomers and in the present invention, all of these possible isomers andmixtures thereof are embraced.

BEST MODE FOR CARRYING OUT THE INVENTION

The 2-pyranone derivatives (I) of the invention can be produced bycombining the N-alkylation reaction and the hydrolytic reaction ofesters. Examples of the N-alkylation reaction include (1) the aminemediated nucleophilic substitution with alkyl halides, alkylsulfonateester or alkyl sulfates and (2) the reductive alkylation betweenaldehydes or ketones and amines in the presence of reducing agents, asdescribed in "SEIMITSU YUKIGOSEI" (Seiichi Takano & Kunio Ogasawara,Nankodo, 1983). Examples of the hydrolytic reaction of esters include 1)hydrolysis using hydrolases such as esterase and lipase and 2)hydrolysis using bases such as sodium hydroxide, sodium hydrogencarbonate and sodium carbonate, as described in Japanese Patent PublicDisclosure No. 2886/1995. More preferably, the reductive alkylation inthe presence of a ketone or aldehyde and a reducing agent or the aminogroup mediated nucleophilic substitution with an alkyl halide may becombined with the hydrolytic reaction with a hydrolase or base to designa production process comprising the following steps A-I:

Step A: Compounds represented by the general formula (III): ##STR3##(where R₇ is an acyl group), either singly or in admixture, aresubjected to reductive alkylation reaction with a ketone or aldehyde ofthe general formula:

    R.sub.1 COR.sub.2 and/or R.sub.3 COR.sub.4

(where R₁, R₂, R₃ and R₄, which may be the same or different, are each ahydrogen atom, an alkyl group, an alkenyl group, an aryl group or anaralkyl group, or an alkyl, alkenyl, aryl or aralkyl group which issubstituted by at least one substituent selected from the groupconsisting of a halogen atom, a hydroxyl group, a lower alkoxy group, alower alkylacyloxy group, a lower alkylacyl group, a loweralkoxycarbonyl group, a nitro group, a cyano group and a heterocyclicgroup, with R₁ and R₂ or R₃ and R₄ being optionally taken together toform an alkylene group, provided that R₁ COR₂ is different from R₃ COR₄)in the presence of a reducing agent to prepare a compound represented bythe general formula (IIa): ##STR4## (where R is a group --NHCHR₁ R₂,--N(CHR₁ R₂)₂ or --N(CHR₁ R₂)CHR₃ R₄ (where R₁, R₂, R₃ and R₄ are thesame as defined above, provided that when R is --N(CHR₁ R₂)CHR₃ R₄, CHR₁R₂ and CHR₃ R₄ are different groups); and R₇ is the same as definedabove).

In compound (III), the acyl group has a carbonyl group bound to a carbonatom in an organic group. In preferred examples of the acyl group, theorganic group is a straight-chained, branched or mono- or polycyclicaliphatic group that have 1-15 carbon atoms, and more preferred examplesinclude butyryl, isobutyryl, isovaleryl, 2-methylbutyryl,4-methylvaleryl, cyclohexanecarbonyl, 4-methylhexanoyl,5-methylhexanoyl, 6-methylheptanoyl, cyclohexylethylcarbonyl, octanoyl,6-methyloctanoyl and 7-methyloctanoyl groups.

Step B: Compound (IIa) is subjected to an ester hydrolysis reactionusing 1) a hydrolase such as porcine liver esterase or lipase or 2) abase such as sodium hydroxide, sodium hydrogencarbonate or sodiumcarbonate to prepare the end compound represented by the general formula(Ia): ##STR5## (where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂ or--N(CHR₁ R₂)CHR₃ R₄ (where R₁, R₂, R₃ and R₄ are the same as definedabove)).

Step C: Compound (III) is hydrolyzed by the same procedure as in step Bto prepare a compound represented by the formula (IV): ##STR6## Step D:Compound (IV) is alkylated reductively by the same procedure as in stepA to prepare the end compound (Ia).

Step E: Compound represented by the general formula (Ia): ##STR7##(where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂ or --N(CHR₁ R₂)CHR₃ R₄(where R₁, R₂, R₃ and R₄ are the same as defined above) is subjected tothe amino group mediated nucleophilic substitution with a compoundrepresented by the general formula R₁ R₂ CHX, R₃ R₄ CHX and/or R₅ R₆ CHX(where R₁, R₂, R₃, R₄, R₅ and R₆, which may be the same or different,are such that R₁, R₂, R₃ and R₄ are the same as defined above and thatR₅ and R₆ are each a hydrogen atom, an alkyl group, an alkenyl group, anaryl group or an aralkyl group, or alkyl, alkenyl, aryl or aralkyl groupwhich is substituted by at least one substituent selected from the groupconsisting of a halogen atom, a hydroxyl group, a lower alkoxy group, alower alkylacyloxy group, a lower alkylacyl group, a loweralkoxycarbonyl group, a nitro group, a cyano group and a heterocyclicgroup, which CHR₅ R₆ optionally forming a cyclic alkyl group, providedthat R₁ R₂ CHX, R₃ R₄ CHX and R₅ R₆ CHX are different from one another;and X is a halogen atom) to prepare the end compound represented by thegeneral formula (Ib): ##STR8## (where R is a group --N⁺ (CHR₁ R₂)₃, --N⁺(CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁ R₂)(CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃,R₄, R₅ and R₆ are the same as defined above)).

Step F: Compound (IV) is subjected to the amino group mediatednucleophilic substitution with a compound represented by the generalformula R₁ R₂ CHX (where R₁ and R₂, which may be the same or different,are each a hydrogen atom, an alkyl group, an alkenyl group, an arylgroup or an aralkyl group, or an alkyl, alkenyl, aryl or aralkyl groupwhich is substituted by at least one substituent selected from the groupconsisting of a halogen atom, a hydroxyl group, a lower alkoxy group, alower alkylacyloxy group, a lower alkylacyl group, a loweralkoxycarbonyl group, a nitro group, a cyano group and a heterocyclicgroup, with CHR₁ R₂ optionally forming a cyclic alkyl group) to preparethe end compound represented by the general formula (Ic): ##STR9##(where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂ or --N⁺ (CHR₁ R₂)₃ (whereR₁ and R₂ are the same as defined above)).

Step G: Compound (IIa) is treated by the same procedure as in step E toprepare a compound represented by the general formula (IIb): ##STR10##(where R is a group --N⁺ (CHR₁ R₂)₃, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺(CHR₁ R₂) (CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃, R₄, R₅ and R₆ are the sameas defined above); and R₇ is the same as defined above).

Step H: Compound (III) is treated by the same procedure as in step F toprepare a compound represented by the general formula (IIc): ##STR11##(where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂ or --N⁺ (CHR₁ R₂)₃ (whereR₁ and R₂ are the same as defined above); and R₇ is the same as definedabove).

Step I: Compound (IIb) is hydrolyzed by the same procedure as in step Bto prepare the end compound (Ib). The same procedure may be employed toprepare the end compound (Ic) from compound (IIc).

It should be mentioned that steps E and G may be carried out withoutisolating compounds (Ia) and (IIa) which are respectively obtained instep D (or B) and step A but subjecting them as such to the associatedreactions.

The above-described production processes of the invention are summarizedin the following charts. ##STR12##

It should be noted that the 2-pyranone derivatives of the generalformula (III) which are used as the starting compound for the productionprocess described above are all known and may be found in J.Antibiotics, 42:1019-1036, 1989, as well as Japanese Patent PublicDisclosure Nos. 304893/1989 and 213758/1993. The 2-pyranone derivativeof the formula (IV) is also a known compound which was initiallyreported to have an antimicrobial activity (Abstracts of the 17th AnnualMeeting of the Pesticide Science Society of Japan, p. 39, 1992) andwhich was recently reviewed for its platelet increasing action (JapanesePatent Public Disclosure No. 2886/1995).

In order to perform the N-alkylation reaction in the above-describedprocess reductively, the starting compound (III) or (IV) may be reactedwith a ketone or aldehyde of the general formula R₁ COR₂ or R₃ COR₄,preferably in an amount of 1-5 moles per mole of the starting compound,in the presence of a reducing agent preferably sodium cyanoborohydride,lithium cyanoborohydride or formic acid, preferably in an amount of0.6-2 moles. Any solvents that dissolve the starting compound withoutinterfering with the reaction may be used without particular limitationsand preferred examples include alcohols such as methanol and ethanol,dimethylformamide and water, which may be used in admixture. Dependingon the case, absolute alcohols or alcoholic solutions of acids such ashydrochloric acid may be added in order to enhance the efficiency of thereaction. The reaction temperature varies with the solvent, reducingagent and the starting material used, etc. and the range of 0-40° C. ispreferred. The reaction time also varies with the solvent, reducingagent and starting material used, etc. and it typically ranges from 30min to 72 hr. After the end of the reaction, the product may berecovered by removing the solvent, adsorbing the residual solution on acolumn such as SepPack C18 (Waters), eluting with a solvent such asmethanol, optionally fractionating by column chromatography as requiredand then freeze-drying the eluate. For reacting the starting compound(III) or (IV) with the ketone or aldehyde of the general formula R₁ COR₂or R₃ COR₄, a two-stage reaction may be adopted in consideration of thereaction affinity.

The amino group mediated nucleophilic substitution can be implemented byreacting a reactant alkyl halide with the starting compound (Ia), (IIa),(III) or (IV) in an inert solvent in the presence or absence of a base.Examples of the base that can be used include metal bases such aspotassium carbonate, sodium carbonate, sodium hydrogen carbonate,potassium hydroxide and sodium hydroxide, and organic bases such astriethylamine, trimethylamine, diisopropylethylamine and pyridine.Examples of the inert solvent that can be used include methanol,ethanol, propanol, tetrahydrofuran and dimethylformamide. The reactiontemperature is preferably 10-100° C. The reaction time which varies withthe solvent, base and the starting materials used, etc. usually rangesfrom 30 min to 3 days. After the end of the reaction, the product may berecovered by removing the solvent from the reaction solution undervacuum in the usual manner and fractionally purifying the residue bycolumn chromatography.

In order to perform the ester hydrolytic reaction in the above-describedprocess by means of a hydrolase, the starting compound (II) or (III) maybe reacted with a hydrolase, preferably selected from (but by no meanslimited to) porcine liver esterase, lipase, acetyl esterase,Takadiastase or cholesterol esterase in a solvent. Preferred examples ofthe solvent that can be used are mixtures of organic solvents such asalcohols (e.g., methanol and ethanol) or ketones (e.g., acetone andmethyl ethyl ketone) with buffer solutions at pH of 6-8. The reactiontemperature which varies with the enzyme used is preferably 10-40° C.The reaction time which varies with the solvent, enzyme and the startingmaterial used, etc. usually ranges from 12 hr to 30 days. After the endof the reaction, the product may be recovered by adsorbing theenzyme-removed reaction mixture on a column such as SepPack C18, elutingwith a solvent such as methanol, further fractionating by columnchromatography as required, and freeze-drying the eluate.

In order to perform the ester hydrolytic reaction by means of a base,the starting compound (II) or (III) may be reacted with a basepreferably selected from (but by no means limited to) alkali metalcarbonates such as sodium carbonate, potassium carbonate, cesiumcarbonate and lithium carbonate, alkali metal hydrogencarbonates such assodium hydrogencarbonate, potassium hydrogencarbonate and lithiumhydrogencarbonate, alkali metal hydroxides such as sodium hydroxide,potassium hydroxide and lithium hydroxide, and alkaline earth metalhydroxides such as barium hydroxide and calcium hydroxide in a solvent.Preferred examples of the solvent that can be used are mixtures oforganic solvents such as alcohols (e.g., methanol and ethanol) andketones (e.g., acetone and methyl ethyl ketone) with water. The reactiontemperature which varies with the base used is preferably 10-40° C. Thereaction time which varies with the solvent, base and the startingmaterial used, etc. usually ranges from 3 hr to 5 days. After the end ofthe reaction, the product may be recovered by removing thewater-miscible organic solvent such as acetone from the reactionsolution under vacuum, performing extraction on the aqueous layer withan organic solvent such as ethyl acetate and fractionally purifying theaqueous layer by column chromatography.

The novel 2-pyranone derivatives of the invention can be used astherapeutics for thrombocytopenia. Therapeutics for thrombocytopenia aredrugs which, when administered into humans, can induce plateletproduction in the body to thereby treat the thrombocytopenia caused byvarious reasons.

The measurement of platelet increasing activity can be implemented bythe method described in Ishibashi, T. et al, Blood, 74(4):1241-1244,1989 or modifications thereof. For example, an animal for plateletmeasurement such as mouse (e.g. C57BL/6 mouse), rat, dog or monkey isadministered intraperitoneally with a test drug as dissolved inethanolic physiological saline or dimethyl sulfoxide (hereundersometimes referred to as DMSO) at a suitable concentration. Thefrequency of administration is usually once or twice a day for 5-10continuous days; a blood sample is taken from the orbital venous plexusseveral hours after the final administration and the platelet count isdetermined. Administration may be performed by any methods includingoral, intravenous, intramuscular and subcutaneous routes. The intervalof administrations, their frequency and the number of days on which theadministration is made are also variable with the drug under test.Platelet count can be determined by an electrical impedance method usinga multi-channel automatic platelet counter (e.g., COULTER COUNTER ModelJT of Coulter Corporation).

The novel 2-pyranone derivatives of the invention may be administered invarious dosage forms, including those for oral administration such astablets, capsules, granules, powders and syrups, and those forparenteral administration such as injections (i.v., i.m. and s.c.),infusions and suppositories. These various pharmaceutical preparationscan be formulated by combining the compounds of the invention withsuitable excipients, binders, disintegrators, lubricants, flavoringagents, coloring agents, solubilizing agents, suspensions, coatingagents, etc.

The compounds of the invention can be administered in doses that arevariable with the symptoms of the disease, the age of the patient, hisbody weight, the method of administration, etc. and the appropriate doseis determined by doctor; usually, the daily dose ranges from 0.01 mg/kgbody weight to 20 mg/kg body weight per adult.

The present invention will now be described in greater detail withreference to the following examples, which are by no means intended tolimit the scope of the invention.

EXAMPLES Example 1

Mouse Platelet Increasing Action

C57BL/6 mice (male, 7-week old) were administered intraperitoneally withthe following compounds of the invention in 1% DMSO physiological salineor with 1% DMSO physiological saline alone (control) at 24-hr intervalsfor 5, 7 or 10 continuous days. Blood samples were collected 72-hr afterthe final administration in the case of 5-day administration, and 4-hrafter the final administration in the case of 7 or 10-dayadministration, and the platelet count was determined by an electricalimpedance method.

6-[3,6-Dihydroxy-3-(2-dimethylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described below in Example 3);

6-[3-(2-Diethylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 7);

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-isopropylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 9);

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-pentylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 12);

6-[3-(2-Cyclopentylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 13);

6-[3-(2-Diethanolaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 20);

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-trimethylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 23);

6-[3-(2-Diallylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one(the compound described in Example 25).

The results obtained are shown in Table 1.

                  TABLE 1                                                         ______________________________________                                                 Dose of   Period of No. of                                                    administ- administ- test   Platelet count,                                    ration,   ration,   animals                                                                              (Mean ± S.E.                           Compound (mg/kg)   (days)    (n)    × 10.sup.3 /μl)                  ______________________________________                                        Control  0         7         8      1070 ± 152                             Compound 0.03      7         6      1417 ± 159                             of EX. 3 0.1       7         7      1486 ± 314                                      1.0       7         8      1532 ± 137                             Control  0         7         8       985 ± 130                             Compound 0.1       10        8      1280 ± 260                             of EX. 3 1.0       10        8      1356 ± 156                             Control  0         5         8      1048 ± 137                             Compound 0.03      5         8      1373 ± 76                              of EX. 7 0.1       5         8      1686 ± 226                                      1.0       5         8      1408 ± 427                             Control  0         5         8      1065 ± 94                              Compound 0.03      5         8      1433 ± 125                             of EX. 9 0.1       5         8      1818 ± 291                                      1.0       5         8      1465 ± 244                             Control  0         5         8      1015 ± 104                             Compound 0.03      5         8      1263 ± 99                              of EX. 12                                                                              0.1       5         8      1623 ± 192                                      1.0       5         8      1338 ± 236                             Control  0         5         8      1015 ± 156                             Compound 0.03      5         8      1490 ± 108                             of EX. 13                                                                              0.1       5         8      1765 ± 291                                      1.0       5         8      1313 ± 293                             Control  0         5         8      1057 ± 146                             Compound 0.03      5         8      1154 ± 138                             of EX. 20                                                                              0.1       5         8      1325 ± 104                                      1.0       5         8      1498 ± 192                             Control  0         5         8      1050 ± 119                             Compound 0.1       5         8      1545 ± 294                             of EX. 23                                                                              1.0       5         8      1513 ± 425                             Control  0         5         8      1000 ± 82                              Compound 0.03      5         8      1074 ± 182                             of EX. 25                                                                              0.1       5         8      1506 ± 209                                      1.0       5         8      1133 ± 170                             ______________________________________                                    

Example 2

Toxicity Test

C57BL/6 mice were administered intravenously with 5 mg/kg of theinvention compound of Example 3 and there were no cases of death.

Example 3

6-[3,6-Dihydroxy-3-(2-dimethylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

Step A:6-[10-(3-Cyclohexylcarbonyloxy)cyclohexyl-3,6-dihydroxy-3-(2-dimethylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-on

A portion (320 mg, 0.5 mmol) of6-[3-(2-aminoethyl)-10-(3-cyclohexylcarbonyloxy)cyclohexyl-3,6-dihydroxy-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-onewas dissolved in 20 ml of methanol and 0.4 ml (5.0 mM) of formalin (35%formaldehyde in aq. sol.) was added. To the resulting solution, 63 mg(1.0 mM) of sodium cyanoborohydride was added under cooling with ice andthen the mixture was stirred at room temperature for 30 min. The solventwas removed under vacuum and the resulting residue was dissolved in 10ml of water, adsorbed on SepPack C18, washed with 100 ml of water andeluted with 50 ml of methanol. The methanol-eluted fraction wasfreeze-dried to give 320 mg of a crude product, which was separated byhigh-performance liquid chromatography (Develosil Packed Column ofNomura Kagaku Co., Ltd.; φ50 mm×300' mm; eluent--the mixture of waterand acetonitrile containing 0.05% trifluoroacetic acid). The peaks ofinterest were combined, diluted with water to 2 times volume, adsorbedon SepPack C18, washed with 100 ml of water and eluted with 50 ml ofmethanol. The methanol-eluted fraction was freeze-dried to give 170 mgof the titled compound (yield=50%).

Mass spectrum (SIMS): m/z=668(M+H)⁺, 690(M+Na)⁺

¹ HNMR(CD₃ OD, δ): 0.95(3H,t,J=7.6 Hz), 1.02-2.03(23H,m), 2.26(2H,m),2.52-2.67(2H,m), 2.86(6H,s), 3.12(1H,m), 3.28(1H,m), 4.31(1H,m),4.69(1H,m), 4.94(1H,m), 5.10(1H,m), 5.31(1H,m), 5.45(1H,m),5.95-6.19(3H,m), 6.27(2H,m), 7.07(1H,m)

Step B:6-[3,6-Dihydroxy-3-(2-dimethylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (50 mg, 0.07 mmol) of the compound synthesized in step A wasdissolved in 18 ml of 0.05 M KH₂ PO₄ -NaOH buffer solution (pH 7.0) and2 ml of methanol; to the solution, 0.5 ml of a suspension of porcineliver derived esterase (30 mg/3 ml) was added and the mixture was shakenovernight at 37° C. The enzyme-removed reaction mixture was adsorbed onSepPack C18, washed with 10 ml of water and eluted with 10 ml ofmethanol. The methanol-eluted fraction was freeze-dried to give 39 mg ofthe titled compound (yield=100%).

Mass spectrum (FAB-MS): m/z=556(M-H)⁻,

¹ HNMR(CD₃ OD, δ): 0.96(3H,t,J=7.2 Hz), 1.02-2.02(14H,m), 2.28(1H,m),2.56(2H,m), 2.85(6H,s), 3.11(1H,m), 3.28(1H,m), 4.31(1H,m), 4.97(1H,m),5.10(1H,m), 5.30(1H,m), 5.43(1H,m), 5.95-6.08(3H,m), 6.25(2H,m),7.08(1H,m)

Step C:6-[3-(2-Aminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (56 mg, 0.09 mmol) of6-[3-(2-aminoethyl)-10-(3-cyclohexylcarbonyloxy)cyclohexyl-3,6-dihydroxy-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-onewas treated by the same procedure as described in step B to prepare 46mg of the titled compound (yield=97%).

Step D:6-[3,6-Dihydroxy-3-(2-dimethylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (50 mg, 0.09 mol) of the compound obtained in step C wastreated by the same procedure as described in step A to prepare 30 mg ofthe titled compound (yield=66%).

Example 4

6-[3-(2-Benzylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (90 mg, 0.17 mmol) of the compound obtained in step C ofExample 3 and 21 mg (0.2 mmol) of benzaldehyde were treated by the sameprocedure as described in step A of Example 3 to prepare 20 mg of thetitled compound (yield=20%).

Mass spectrum (FAB-MS): m/z=618(M-H)⁻,

¹ HNMR(CD₃ OD, δ): 0.94(3H,t,J=7.2Hz), 1.02-2.00(13H,m), 2.26(1H,m),2.55(2H,m), 3.08(1H,m), 3.17(1H,m), 3.54(1H,m), 4.18(2H,m), 4.31(1H,m),4.95(1H,m), 5.07(1H,m), 5.31(1H,m), 5.44(1H,m), 5.92-6.03(3H,m),6.26(2H,m), 7.09(1H,m), 7.47(5H,m)

Example 5

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-octylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (90 mg, 0.17 mmol) of the compound obtained in step C ofExample 3 and 26 mg (0.2 mmol) of octyl aldehyde were treated by thesame procedure as described in step A of Example 3 to prepare 50 mg ofthe titled compound (yield=46%).

Mass spectrum (FAB-MS): m/z=640(M-H)⁻

¹ HNMR(CD₃ OD, δ): 0.94(3H,m), 0.98(3H,m), 1.02-2.00(25H,m), 2.21(1H,m),2.56(2H,m), 2.96(2H,m), 3.04(1H,m), 3.12(1H,m), 3.54(1H,m), 4.31(1H,m),4.95(1H,m), 5.10(1H,m), 5.30(1H,m), 5.43(1H,m), 5.94-6.08(3H,m),6.26(2H,m), 7.09(1H,m)

Example 6

Pharmaceutical Formulation

The compound (4 g) obtained in Example 3 and mannitol (50 g) weredissolved in water for injection (100 ml) containing 30% (w/w) ofpolyethylene glycol 400 and the solution was sterilized by filtrationmethod. The filtered solution was dispensed in 1 ml portions intoampules to prepare injections.

Example 7

6-[3-(2-Diethylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 40 μl (0.73 mmol) of acetaldehyde were treated by the sameprocedure as described in step A of Example 3 to prepare 35 mg of thetitled compound (yield=20%).

Mass spectrum (FAB-MS): m/z=586(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.97(3H,t,J=7.4 Hz), 1.00-2.00(19H,m), 2.27(1H,m),2.55(2H,m), 3.07(1H,m), 3.21(5H,m), 3.54(1H,m), 4.31(1H,m), 4.94(1H,m),5.11(1H,m), 5.31(1H,m), 5.44(1H,m), 5.97-6.03(3H,m), 6.24(2H,m),7.08(1H,m)

Example 8

6-[3,6-Dihydroxy-3-(2-(dipropylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 65 μl (0.91 mmol) of propionaldehyde were treated by thesame procedure as described in step A of Example 3 to prepare 76 mg ofthe titled compound (yield=41%).

Mass spectrum (FAB-MS): m/z=614(M+H)⁺

¹ HNMR(CD₃ OD, δ): 1.02(9H,m), 1.10-2.00(17H,m), 2.31(1H,m), 2.56(2H,m),3.09(1H,m), 3.30(5H,m), 3.54(1H,m), 4.30(1H,m), 4.94(1H,m), 5.10(1H,m),5.31(1H,m), 5.44(1H,m), 5.97-6.03(3H,m), 6.25(2H,m), 7.07(1H,m)

Example 9

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-isopropylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 27 μl (0.36 mmol) of acetone were treated by the sameprocedure as described in step A of Example 3 to prepare 40 mg of thetitled compound (yield=23%).

Mass spectrum (FAB-MS): m/z=572(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.96(3H,t,J=7.4 Hz), 1.00-2.00(20H,m), 2.22(1H,m),2.56(2H,m), 3.07(1H,m), 3.13(1H,m), 3.55(1H,m), 4.31(1H,m), 4.94(1H,m),5.09(1H,m), 5.31(1H,m), 5.44(1H,m), 5.90-6.03(3H,m), 6.25(2H,m),7.08(1H,m)

Example 10

6-[3,6-Dihydroxy-3-(2-dipentylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExmaple 3 and 96 μl (0.91 mmol) of valeryl aldehyde were treated by thesame procedure as described in step A of Example 3 to prepare 53 mg ofthe titled compound (yield=26%).

Mass spectrum (FAB-MS): m/z=670(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(9H,m), 1.00-2.10(25H,m), 2.25(1H,m), 2.56(1H,m),3.09(6H,m), 3.54(1H,m), 4.30(1H,m), 4.95(1H,m), 5.11(1H,m) 5.31(1H,m),5.44(1H,m), 5.90-6.10(3H,m), 6.25(2H,m), 7.08(1H,m)

Example 11

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-methyloctylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 52 μl (0.33 mmol) of octyl aldehyde were N-octylated bythe same procedure as described in step A of Example 3; without beingsubsequently isolated, the product was N-methylated with 35%formaldehyde solution (72 ml, 0.90 mmol) to prepare 70 mg of the titledcompound (yield=35%).

Mass spectrum (FAB-MS): m/z=656(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.94(3H,m), 0.99(3H,m), 1.00-2.20(25H,m), 2.28(1H,m),2.56(2H,m), 2.82(3H,s), 3.09(4H,m), 3.54(1H,m), 4.31(1H,m), 4.95(1H,m),5.10(1H,m), 5.31(1H,m), 5.44(1H,m), 6.00-6.20(3H,m), 6.25(2H,m),7.08(1H,m)

Example 12

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-pentylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 38 μl (0.36 mmol) of valeryl aldehyde were treated by thesame procedure as described in step A of Exmaple 3 to prepare 84 mg ofthe titled compound (yield=46%).

Mass spectrum (FAB-MS): m/z=600(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(6H,m), 1.05-2.10(19H,m), 2.22(1H,m), 2.56(2H,m),2.97(2H,m), 3.09(1H,m), 3.15(1H,m), 3.54(1H,m), 4.31(1H,m), 4.94(1H,m),5.10(1H,m), 5.31(1H,m), 5.44(1H,m), 5.90-6.20(3H,m), 6.25(2H,m),7.08(1H,m)

Example 13

6-[3-(2-Cyclopentylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExmaple 3 and 32 μl (0.36 mmol) of cyclopentanone were treated by thesame procedure as described in step A of Example 3 to prepare 152 mg ofthe titled compound (yield=84%).

Mass spectrum (FAB-MS): m/z=598(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.96(3H,m), 1.00-2.10(21H,m), 2.21(1H,m), 2.57(2H,m),3.05(1H,m), 3.14(1H,m), 3.52(2H,m), 4.30(1H,m), 4.89(1H,m), 5.11(1H,m),5.31(1H,m), 5.44(1H,m), 5.90-6.20(3H,m), 6.27(2H,m), 7.10(1H,m)

Example 14

6-[3-(2-Cyclohexylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 38 μl (0.36 mmol) of cyclohexanone were treated by thesame procedure as described in step A of Example 3 to prepare 146 mg ofthe titled compound (yield=79%).

Mass spectrum (FAB-MS): m/z=612(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.96(3H,t,J=7.4 Hz), 1.00-2.10(23H,m), 2.21(1H,m),2.56(2H,m), 3.08(2H,m), 3.15(1H,m), 3.54(1H,m), 4.30(1H,m), 4.97(1H,m),5.11(1H,m), 5.31(1H,m), 5.43(1H,m), 5.90-6.20(3H,m), 6.27(2H,m),7.11(1H,m)

Example 15

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-methylpentylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (100 mg, 0.17 mmol) of the compound obtained in Example 12 and20 μl (0.25 mmol) of 35% formaldehyde solution were treated by the sameprocedure as described in step A of Example 3 to prepare 80 mg of thetitled compound (yield 78%).

Mass spectrum (FAB-MS): m/z=614(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.96(6H,m), 1.00-2.10(19H,m), 2.28(1H,m), 2.57(2H,m),2.83(3H,s), 3.09(3H,m), 3.31(1H,m), 3.55(1H,m), 4.31(1H,m), 4.95(1H,m),5.10(1H,m), 5.32(1H,m), 5.43(1H,m), 6.00-6.20(3H,m), 6.27(2H,m),7.09(1H,m)

Example 16

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-pentan-3-yl-aminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in Example 12 and37 μl (0.36 mmol) of 3-pentanone were treated by the same procedure asdescribed in step A of Example 3 to prepare 141 mg of the titledcompound (yield=78%).

Mass spectrum (FAB-MS): m/z=600(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.98(9H,m), 1.00-2.10(17H,m), 2.25(1H,m), 2.56(2H,m),3.02(1H,m), 3.12(2H,m), 3.54(1H,m), 4.29(1H,m), 4.96(1H,m), 5.11(1H,m),5.31(1H,m), 5.43(1H,m), 5.90-6.20(3H,m), 6.27(2H,m), 7.08(1H,m)

Example 17

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-methyl-isopropylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (114 mg, 0.20 mmol) of the compound obtained in Example 9 and50 μl (0.60 mmol) of 35% formaldehyde solution were treated by the sameprocedure as described in step A of Example 3 to prepare 95 mg of thetitled compound (yield =80%).

Mass spectrum (FAB-MS): m/z=586(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(3H,t,J=7.4 Hz), 1.00-1.81(17H,m),1.24(3H,d,J=6.6 Hz), 1.26(3H,d,J=6.6 Hz), 2.41(2H,m), 2.54(2H,m),2.67(3H,s), 3.02(1H,m), 3.16(1H,m), 3.51(2H,m), 4.27(1H,m), 5.02(1H,m),5.08(1H,m), 5.27(1H,m), 5.44(1H,m), 5.94-6.13(3H,m), 6.25(2H,m),7.06(1H,m)

Example 18

6-[3-(2-Dibutylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (130 mg, 0.25 mmol) of the compound obtained in step C ofExample 3 and 72 mg (1.0 mmol) of butyl aldehyde were treated by thesame procedure as described in step A of Example 3 to prepare 80 mg ofthe titled compound (yield 50%).

Mass spectrum (FAB-MS): m/z=642(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95-1.02(9H,m), 1.12-2.00(28H,m), 2.29(1H,m),2.56(2H,m), 3.11(4H,m), 3.54(1H,m), 4.30(1H,m), 4.96(1H,m), 5.11(1H,m),5.31(1H,m), 5.44(1H,m), 5.97-6.10(3H,m), 6.25(2H,m), 7.08(1H,m)

Example 19

6-[3-[2-(1,3-Dihydroxyisopropylaminoethyl)]-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (53 mg, 0.10 mmol) of the compound obtained in step C ofExample 3 and 22 mg (0.12 mmol) of dihydroxyacetone (dimer) were treatedby the same procedure as described in step A of Example 3 to prepare 50mg of the titled compound (yield=80%).

Mass spectrum (FAB-MS): m/z=604(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.97(3H,t,J=7.4 Hz), 1.00-2.00(13H,m), 2.34(1H,m),2.56(2H,m), 3.25(2H,m), 3.37(1H,m), 3.55(1H,m), 3.72(2H,m), 3.82(2H,m),4.31(1H,m), 4.97(1H,m), 5.11(1H,m), 5.31(1H,m), 5.45(1H,m),5.94-6.14(3H,m), 6.27(2H,m), 7.09(1H,m)

Example 20

6-[3-(2-Diethanolaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (160 mg, 0.30 mmol) of the compound obtained in step C ofExample 3 and 87 mg (0.73 mmol) of hydroxyacetaldehyde were treated bythe same procedure as described in step A of Example 3 to prepare 182 mgof the titled compound (yield=98%).

Mass spectrum (FAB-MS): m/z=618(M+H)⁺

¹ HNMR(CD₃ OD, δ): 1.96(3H,t,J=7.4 Hz), 1.10-1.94(14H,m), 2.35(1H,m),2.55(2H,m), 2.96(2H,m), 3.09(2H,m), 3.19(1H,m), 3.54(1H,m), 3.79(4H,m),4.30(1H,m), 5.01(1H,m), 5.09(1H,m), 5.29(1H,m), 5.44(1H,m),5.94-6.13(3H,m), 6.25(2H,m), 7.07(1H,m)

Example 21

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-[2-(1-pyridiniumisopropylaminoethyl)]-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (62 mg, 0.12 mmol) of the compound obtained in step C ofExample 3 and 87 mg (0.51 mmol) of 1-acetonylpyridinium chloride weretreated by the same procedure as described in step A of Example 3 toprepare 30 mg of the titled compound (yield=39%).

Mass spectrum (FAB-MS): m/z=650(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.93(3H,m), 1.16(3H,m), 1.00-1.91(14H,m), 2.53(2H,m),2.60-2.91(3H,m), 3.47(1H,m), 4.16(1H,m), 4.41(1H,m), 4.64(1H,m),4.92(1H,m), 5.04(1H,m), 5.29(1H,m), 5.41(1H,m), 5.91(2H,m), 6.00(1H,m),6.24(2H,m) 7.07(1H,m), 8.12(2H,m), 8.59(1H,m), 8.93(2H,m)

Example 22

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-trimethylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (190 mg, 0.34 mmol) of the compound obtained in Exmaple 3 and43 μl (0.68 mmol) of methyl iodide were dissolved in 2 ml ofdimethylformamide and stirred at room temperature for 1.5 hr. Thesolvent was removed under vacuum to give a crude product, which wasseparated by high-performance liquid chromatography. The peaks ofinterest were combined, deacidified on SepPack C18 and freeze-dried toprepare 80 mg of the titled compound (yield=41%).

Mass spectrum (FAB-MS): m/z=572(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(3H,m), 1.00-1.98(14H,m), 2.25(1H,m), 2.56(2H,m),3.10(9H,s), 3.54(2H,m), 4.25(1H,m), 5.08(2H,m), 5.26(1H,m), 5.42(1H,m),5.90-6.10(3H,m), 6.25(2H,m) 7.08(1H,m),

Example 23

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-trimethylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (62 mg, 0.12 mmol) of the compound obtained in step C ofExample 3 and 46 μl (0.84 mmol) of methyl iodide were treated by thesame procedure as described in Example 22 to prepare 40 mg of the titledcompound (yield=58%).

Example 24

6-[3-(2-Allylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

A portion (87 mg, 0.16 mmol) of the compound obtained in step C ofExample 3, 111 μl (1.28 mmol) of allyl bromide and 280 μl (1.60 mmol) ofdiisopropylethylamine were dissolved in 8 ml of methanol and stirred at50° C. for 4 hr. The solvent was removed under vacuum to give a crudeproduct, which was separated by high-performance liquid chromatography.The peaks of interest were combined, deacidified on SepPack C18 andfreeze-dried to prepare 6 mg of the titled compound (yield=7%).

Mass spectrum (FAB-MS): m/z=570(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(3H,m), 1.05-1.95(13H,m), 2.26(1H,m), 2.56(2H,m),3.06(1H,m), 3.16(1H,m), 3.15(1H,m), 3.55(1H,m), 3.63(2H,m), 4.31(1H,m),4.94(1H,m), 5.10(1H,m), 5.31(1H,m), 5.42-5.54(3H,m), 5.87-6.09(4H,m)6.27(2H,m), 7.08(1H,m),

Example 25

6-[3-(2-Diallylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

The same procedure as described in Example 24 was followed to prepare 22mg of the titled compound (yield=23%).

Mass spectrum (FAB-MS): m/z=610(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.95(3H,m), 1.05-1.95(14H,m), 2.06(1H,m), 2.30(1H,m),2.56(2H,m), 3.06(1H,m), 3.55(1H,m), 3.63-3.81(4H,m), 4.31(1H,m),4.94(1H,m), 5.10(1H,m), 5.31(1H,m), 5.44(1H,m), 5.57-5.61(4H,m)5.91-6.06(4H,m), 6.27(2H,m) 7.08(1H,m)

Example 26

6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-triallylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one

The same procedure as described in Example 24 was followed to prepare 20mg of the titled compound (yield=19%).

Mass spectrum (FAB-MS): m/z=651(M+H)⁺

¹ HNMR(CD₃ OD, δ): 0.96(3H,m), 1.05-1.95(12H,m), 2.18-2.35(2H,m),2.57(2H,m), 3.13(1H,m), 3.41(1H,m), 3.56(1H,m), 3.93(6H,m), 4.28(1H,m),4.95(1H,m), 5.09(1H,m), 5.31(1H,m), 5.44(1H,m), 5.71-5.77(6H,m),5.99-6.08(6H,m), 6.25(2H,m) 7.08(1H,m)

We claim:
 1. A compound represented by the general formula (I):##STR13## (where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂, --N(CHR₁R₂)CHR₃ R₄, --N⁺ (CHR₁ R₂)₃, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁R₂)(CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃, R₄, R₅ and R₆, which may be thesame or different, are each a hydrogen atom, an alkyl group, an alkenylgroup, an aryl group or an aralkyl group, or an alkyl, alkenyl, aryl oraralkyl group which is substituted by at least one substituent selectedfrom the group consisting of a halogen atom, a hydroxyl group, a loweralkoxy group, a lower alkylacyloxy group, a lower alkylacyl group, alower alkoxycarbonyl group, a nitro group, and a cyano group, with CHR₁R₂, CHR₃ R₄ or CHR₅ R₆ optionally forming a cyclic alkyl group, providedthat R is not --NHCH₃, and that when R is --N(CHR₁ R₂)CHR₃ R₄, --N⁺(CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁ R₂)(CHR₃ R₄)CHR₅ R₆, CHR₁ R₂, CHR₃ R₄and CHR₅ R₆ are different groups)) or a pharmacologically acceptablesalt thereof.
 2. A compound represented by the general formula (II):##STR14## (where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂, --N(CHR₁R₂)CHR₃ R₄, --N⁺ (CHR₁ R₂)₃, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁R₂)(CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃, R₄, R₅ and R₆, which may be thesame or different, are each a hydrogen atom, an alkyl group, an alkenylgroup, an aryl group or an aralkyl group, or an alkyl, alkenyl, aryl oraralkyl group which is substituted by at least one substituent selectedfrom the group consisting of a halogen atom, a hydroxyl group, a loweralkoxy group, a lower alkylacyloxy group, a lower alkylacyl group, alower alkoxycarbonyl group, a nitro group, and a cyano group, with CHR₁R₂, CHR₃ R₄ or CHR₅ R₆ optionally forming a cyclic alkyl group, providedthat R is not --NHCH₃, and that when R is --N(CHR₁ R₂)CHR₃ R₄, --N⁺(CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁ R₂)(CHR₃ R₄)CHR₅ R₆, CHR₁ R₂, CHR₃ R₄and CHR₅ R₆ are different groups); and R₇ is an acyl group, or a saltthereof.
 3. A compound represented by the general formula (II):##STR15## (where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂, --N(CHR₁R₂)CHR₃ R₄, --N⁺ (CHR₁ R₂)₃, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁R₂)(CHR₃ R₄)CHR₅ R₆ (where R₁, R₂, R₃, R₄, R₅ and R₆, which may be thesame or different, are each a hydrogen atom, an alkyl group, an alkenylgroup, an aryl group or an aralkyl group, or an alkyl, alkenyl, aryl oraralkyl group which is substituted by at least one substituent selectedfrom the group consisting of a halogen atom, a hydroxyl group, a loweralkoxy group, a lower alkylacyloxy group, a lower alkylacyl group, alower alkoxycarbonyl group, a nitro group, a cyano group and aheterocyclic group, with CHR₁ R₂, CHR₃ R₄ or CHR₅ R₆ optionally forminga cyclic alkyl group, provided that R is not --NHCH₃, and that when R is--N(CHR₁ R₂)CHR₃ R₄, --N⁺ (CHR₁ R₂)₂ CHR₃ R₄ or --N⁺ (CHR₁ R₂)(CHR₃R₄)CHR₅ R₆, CHR₁ R₂, CHR₃ R₄ and CHR₅ R₆ are different groups); and R₇is a butyryl group, an isobutyryl group, an isovaleryl group, a2-methylbutyryl group, a 4-methylvaleryl group, a cyclohexanecarbonylgroup, a 4-methylhexanoyl group, a 5-methylhexanoyl group, a6-methylheptanoyl group, a cyclohexylethylcarbonyl group, an octanoylgroup, a 6-methyloctanoyl group or a 7-methyloctanoyl group) or a saltthereof.
 4. A compound according to claim 1, wherein R in the generalformula (I) is --NHCHR₁ R₂, provided that R₁ is not a hydrogen atom whenR₂ is a hydrogen atom, or a pharmacologically acceptable salt thereof.5. A compound according to claim 1, wherein R in the general formula (I)is --N(CHR₁ R₂)CHR₃ R₄, or a pharmacologically acceptable salt thereof.6. A compound according to claim 1, wherein R in the general formula (I)is --N(CHR₁ R₂)₂ or a pharmacologically acceptable salt thereof.
 7. Acompound according to claim 1, wherein R in the general formula (I) is--N⁺ (CHR₁ R₂)₃ or a pharmacologically acceptable salt thereof.
 8. Acompound according to claim 1, wherein R in the general formula (I) is--N⁺ (CHR₁ R₂)₂ CHR₃ R₄, or a pharmacologically acceptable salt thereof.9. A compound according to claim 1, wherein R in the general formula (I)is --N⁺ (CHR₁ R₂)(CHR₃ R₄)CHR₅ R₆, or a pharmacologically acceptablesalt thereof.
 10. A compound according to claim 1, which is selectedfrom the followingcompounds:6-[3,6-Dihydroxy-3-(2-dimethylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3-(2-Diethylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-3-(2-dipropylaminoethyl)-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-isopropylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-pentylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3-(2-Cyclopentylaminoethyl)-3,6-dihydroxy-10-(3-(hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3-(2-Cyclohexylaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-pentan-3-yl-aminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-3-(2-methyl-isopropylaminoethyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3-[2-(1,3-Dihydroxyisopropylaminoethyl)]-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3-(2-Diethanolaminoethyl)-3,6-dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-trimethylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one;6-[3,6-Dihydroxy-10-(3-hydroxycyclohexyl)-4-(phosphonoxy)-3-(2-triallylammoniumethyl)-1,7,9-decatrienyl]-5,6-dihydro-5-ethyl-2H-pyran-2-one,or a pharmacologically acceptable salt thereof.
 11. A Process whichcomprises subjecting compounds, either singly or in admixture, that arerepresented by the general formula (III): ##STR16## (where R₇ is an acylgroup) to reductive alkylation reaction with a ketone or aldehyde of thegeneral formula R₁ COR₂ and/or R₃ COR₄ (where R₁, R₂, R₃ and R₄, whichmay be the same or different, are each a hydrogen atom, an alkyl group,an alkenyl group, an aryl group or an aralkyl group, or an alkyl,alkenyl, aryl or aralkyl group which is substituted by at least onesubstituent selected from the group consisting of a halogen atom, ahydroxyl group, a lower alkoxy group, a lower alkylacyloxy group, alower alkylacyl group, a lower alkoxycarbonyl group, a nitro group, anda cyano group, with R₁ and R₂ or R₃ and R₄ being optionally takentogether to form an alkylene group, provided that R₁ COR₂ is differentfrom R₃ COR₄, and that R₁ is not a hydrogen atom when R₂ is a hydrogenatom) in the presence of a reducing agent, to thereby produce a compoundrepresented by the general formula (IIa) or a salt thereof: ##STR17##(where R is a group --NHCHR₁ R₂, --N(CHR₁ R₂)₂ or --N(CHR₁ R₂)CHR₃ R₄(where R₁, R₂, R₃ and R₄ are the same as defined above)).
 12. A processwhich comprises subjecting a compound represented by the general formula(IV): ##STR18## to reductive alkylation reaction with a ketone oraldehyde of the general formula R₁ COR₂ and/or R₃ COR₄ (where R₁, R₂,R₃, and R₄, which may be the same or different, are each a hydrogenatom, an alkyl group, an alkenyl group, an aryl group or an aralkylgroup, or an alkyl, alkenyl, aryl or aralkyl group which is substitutedby at least one substituent selected from the group consisting of ahalogen atom, a hydroxyl group, a lower alkoxy group, a loweralkylacyloxy group, a lower alkylacyl group, a lower alkoxycarbonylgroup, a nitro group, and a cyano group, with R₁ and R₂ or R₃ and R₄being optionally taken together to form an alkylene group, provided thatR₁ COR₂ is different from R₃ COR₄, and that R₁ is not a hydrogen atomwhen R₂ is a hydrogen atom) in the presence of a reducing agent, tothereby prepare a compound represented by the general formula (Ia) or apharmacologically acceptable salt thereof: ##STR19## (where R is a group--NHCHR₁ R₂, --N(CHR₁ R₂)₂ or --N(CHR₁ R₂)CHR₃ R₄ (where R₁, R₂, R₃ andR₄ are the same as defined above)).
 13. A therapeutic agent forthrombocytopenia containing as an effective ingredient the compound ofclaim 1 or a pharmacologically acceptable salt thereof.